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. 2013 May 27;201(5):693–707. doi: 10.1083/jcb.201302145

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© 2013 Reczek et al.

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Figure 2. — The Ctip–S326A (SA) polypeptide does not interact with Brca1 in vivo. (A) Nuclear extracts of Ctip^+/+, Ctip^S326A/+, and Ctip^S326A/S326A MEFs were fractionated by SDS-PAGE and immunoblotted with antibodies specific for α-tubulin, Brca1, and Ctip. A nonspecific band in the Brca1 immunoblot is designated as “ns”. (B) To evaluate the Brca1–Ctip interaction, the extracts were immunoprecipitated with mouse Brca1-specific antiserum (B1) or the corresponding preimmune serum (Pre) and immunoblotted with Ctip-specific monoclonal antibodies. As shown, Ctip was coimmunoprecipitated with Brca1 from Ctip^+/+ and Ctip^S326A/+ cells, but not Ctip^S326A/S326A cells. Note: the amount of nuclear extract used for immunoblotting (A) represents 6.25% of the total extract used for coimmunoprecipitation analysis (B).